1H NMR detection of vitamin C in human brain in vivo.

Vitamin C (ascorbate) is well established as an essential nutrient that functions as an antioxidant. Since it is present in the human brain at detectable concentrations, this study was designed to detect and quantify ascorbate in the human brain in vivo using 1H NMR spectroscopy (MRS). Ascorbate was consistently detected in all five study subjects, and was measured using MEGA-PRESS difference editing. The in vivo resonance pattern was consistent with that of ascorbate based on position, line width, peak pattern, and relative intensity. Metabolites with a potential for coediting were assessed using phantom solutions. The putative resonances of myo-inositol, lactate, glycerophosphocholine, phosphocholine, and phosphoethanolamine were detected at positions distinct from those of ascorbate. This study represents the first in vivo detection of vitamin C in the human brain using 1H MRS. A concentration of 1.3 +/- 0.3 micromol/g (mean +/- SD, N = 4) was estimated.     

Volume-selective proton MR spectroscopy for in-vitro quantification of anticonvulsants

Administration of anticonvulsant drugs is clinically monitored by checking seizure frequency and by determining the serum concentration of the drug. In a few reports, drug concentrations in brain parenchyma have been determined using ex vivo techniques. Little is known about the in vivo concentration in the brain parenchyma. Our goals were to characterise the NMR spectra of the anticonvulsants at therapeutic concentrations, to determine the minimum detectable concentrations, and to quantify the drugs noninvasively. Volume-selective 1H-MR spectroscopy (MRS) was performed under standard clinical conditions using a single-voxel STEAM (stimulated-echo acquisition mode) sequence at 1.5 T. Spectra of the anticonvulsants carbamazepine, phenobarbital, phenytoin and valproate were acquired in vitro in hydrous solutions at increasing dilution. Phenytoin, phenobarbital and valproate were detectable below maximum therapeutic serum concentrations. Within therapeutic ranges, there was good agreement between concentrations determined by 1H-MRS and those by standard fluorescence polarisation immunoassay. Due to the absence of signals of brain metabolites, the aromatic protons of phenobarbital, phenytoin and carbamazepine, with resonance lines around 7.4 ppm, allow the drugs to be detected. Valproate, with two resonances around 1.2 ppm, should be differentiable from potential brain metabolites using nonlinear analysis of the brain spectrum. Volume-selective 1H-MRS is therefore expected to be able to monitor anticonvulsant therapy in vivo. Received: 14 August 2000 Accepted: 8 September 2000     

Detection of an antioxidant profile in the human brain in vivo via double editing with MEGA-PRESS.

Vitamin C (ascorbate) and glutathione (GSH) are the two most concentrated non-enzymatic antioxidants in the human brain. Double editing with (DEW) MEGA-PRESS at 4T was designed in this study to measure both antioxidants in the same amount of time previously required to measure one. In the occipital lobe of four human subjects, resolved ascorbate (Asc) and GSH resonances were detected repeatedly and simultaneously using DEW MEGA-PRESS. The Asc and GSH concentrations measured using LCModel analysis of DEW MEGA-PRESS spectra were 0.8 +/- 0.1 and 1.0 +/- 0.1 micromol/g (mean +/- SD), with average Cramer-Rao lower bounds (CRLB) of 10% and 7%, respectively. Aside from the effects of J-modulation at a common echo time (TE), double editing did not compromise sensitivity. To determine the extent to which the oxidized forms of Asc and GSH contribute to DEW MEGA-PRESS spectra in vivo, chemical shifts and coupling constants for dehydroascorbate (DHA) and oxidized glutathione (GSSG) were measured at physiologic pH and temperature. DHA does not contribute to the 3.73 ppm DEW MEGA-PRESS Asc resonance. GSSG contributions to the DEW MEGA-PRESS GSH resonance (3.0 ppm) are negligible under physiologic conditions, and would be evidenced by a distinct GSSG resonance (3.3 ppm) at exceptionally high concentrations.     

Quantitative analysis of the effects of physiologic brain motion on point-resolved spectroscopy

BACKGROUND AND PURPOSE: Although single-voxel proton MR spectroscopy is a noninvasive method that enables measurement of brain metabolite concentrations, it has been shown that physiologic brain motion causes inaccuracies in measurement of metabolite concentrations and increases the overall SD of the measurements when the stimulated echo acquisition mode (STEAM) is used. We tested the hypothesis that the point-resolved spectroscopy (PRESS) technique is less sensitive to physiologic brain motion than the STEAM technique. METHODS: In 10 healthy subjects, spectra were obtained from a voxel located in the left basal ganglia by using the PRESS sequence with cardiac gating and without water suppression to assess global phase change as a function of physiologic brain motion. This was accomplished by acquiring data at various time delays from the R wave throughout the cardiac cycle. Subsequently, spectra were obtained in 10 healthy subjects by using PRESS both without and with cardiac gating, and with water suppression, to determine whether brain motion resulted in a statistically significant difference in mean and SD of measured metabolite concentration. RESULTS: At various time delays from the R wave throughout the cardiac cycle, no significant global phase difference was noted in water signal intensity. In addition, when PRESS data were obtained both without and with cardiac gating (by using an optimal delay obtained from previously published data by using STEAM), no significant difference was seen in measured metabolite concentrations and SDs. CONCLUSION: The PRESS technique is relatively insensitive to physiologic brain motion.     

Localized proton NMR spectroscopy in different regions of the human brain in vivo. Relaxation times and concentrations of cerebral metabolites

High-resolution proton NMR spectra of normal human brain in vivo have been obtained from selected 27- and 64-ml volumes-of-interest (VOI) localized in the insular area, the occipital area, the thalamus, and the cerebellum of normal volunteers. Localization was achieved by stimulated echo (STEAM) sequences using a conventional 1.5-T whole-body MRI system (Siemens Magnetom). The proton NMR spectra show resonances from lipids, lactate, acetate, N-acetylaspartate (NAA), gamma-aminobutyrate, glutamine, glutamate, aspartate, creatine and phosphocreatine, choline-containing compounds, taurine, and inositols. While T1 relaxation times of most of these metabolites were about 1100-1700 ms without significant regional differences, their T2 relaxation times varied between 100 and 500 ms. The longest T2 values of about (500 +/- 50) ms were observed for the methyl protons of NAA in the white matter of the occipital lobe compared to (320 +/- 30) ms in the other parts of the brain. No significant regional T2 differences were found for choline and creatine methyl resonances. The relative concentrations of NAA in gray and white matter were found to be 35% higher than those in the thalamus and cerebellum. Assuming a concentration of 10 mM for total creatine the resulting NAA concentrations of 13-18 mM are by a factor of 2-3 higher than previously reported using analytical techniques. Cerebral lactate reached a maximum concentration of about 1.0 mM.     

Short‐echo,single‐shot,full‐intensity proton magnetic resonance spectroscopy for neurochemical profiling at 4 T: Validation in the cerebellum and brainstem

A short echo time (TE = 24 ms) semiadiabatic localization by adiabatic selective refocusing (LASER) sequence was designed and optimized for full‐intensity proton magnetic resonance spectroscopy (¹H MRS) at 4 T. The sequence was combined with VAPOR water suppression and three‐dimensional outer volume suppression for improved localization and suppression of unwanted coherences. Artifact‐free, single‐shot spectra were obtained from the human brain with a spectral pattern almost identical to that obtained with an ultra‐short TE (TE = 5 ms) stimulated‐echo acquisition mode (STEAM) sequence as a result of the train of adiabatic refocusing pulses in semi‐LASER that reduce the apparent TE. Approximately 2‐fold higher signal intensity relative to STEAM was demonstrated in phantoms and the human brain. To test the performance of the sequence in clinically relevant brain regions with a volume coil, semi‐LASER spectra were acquired from three cerebellar and brainstem volumes of interest (VOIs) in 23 healthy subjects. Ultra‐short echo STEAM spectra were acquired from the same VOIs to compare neurochemical profiles obtained with semi‐LASER with those obtained with STEAM. Neurochemical profiles of the cerebellum and brainstem acquired by these two techniques were nearly identical, validating the accuracy of the metabolite concentrations obtained with semi‐LASER at the longer TE relative to STEAM. A high correlation between metabolite concentrations obtained by these two proton ¹H MRS techniques indicated the sensitivity to detect intersubject variation in metabolite levels. Magn Reson Med, 2010. © 2010 Wiley‐Liss, Inc.     

Developmental and regional changes in the neurochemical profile of the rat brain determined by in vivo 1H NMR spectroscopy.

Sixteen metabolites were quantified from 11-24 micro l volumes in three different brain regions (hippocampus, striatum, and cerebral cortex) during postnatal development. Rat pups from the same litter were repeatedly measured on postnatal days 7, 10, 14, 21, and 28 using a completely noninvasive and longitudinal study design. Metabolite quantification was based on ultra-short echo-time (1)H NMR spectroscopy at 9.4 T and LCModel processing. Most of the brain metabolites were quantified with Cramer-Rao lower bounds (CRLB) less than 20%, which corresponded to an estimated concentration error <0.2 micro mol/g. Taurine and total creatine were quantified with CRLB < or = 5% from all 114 processed spectra. The resulting high reliability and reproducibility revealed significant regional and age-related changes in metabolite concentrations. The most sensitive markers for developmental and regional variations between hippocampus, striatum, and cerebral cortex were N-acetylaspartate, myo-inositol, taurine, glutamate, and choline compounds. Absolute values of metabolite concentrations were in very good agreement with previously published in vitro results based on chromatographic measurements of brain extracts. The current data may serve as a reference for studies focused on developmental defects and pathologies using neonatal rat models.     

A homonuclear shift correlated and spatially localized spectroscopy using stimulated echoes

A new method which combines localized high-resolution proton NMR spectroscopy with two-dimensional correlated spectroscopy using stimulated echoes is presented. Stimulated-echo correlated spectroscopy (STECSY) is a straightforward extension of the STEAM method. Experiments with phantoms illustrate the efficacy of STECSY. An in situ application on rat adipose tissue demonstrates that STECSY is a helpful tool with which to select and assign resonances in complex 1H NMR spectra.     

Proton MR spectroscopic characteristics of pediatric pilocytic astrocytomas.

PURPOSEWe report the common characteristics of juvenile pilocytic astrocytomas revealed by proton MR spectroscopy.METHODSEight children with pilocytic astrocytomas were studied with proton MR spectroscopy. The selected sampling volume was approximately 4 cm3, obtained from solid tumor. To localize the sampling volume, we used point-resolved spectroscopy (PRESS) and stimulated-echo acquisition mode (STEAM) techniques to acquire long- and short-TE spectra, respectively. Spectra from PRESS and STEAM sequences were processed using Lorentzian-to-Gaussian transformation and exponential apodization, respectively. For PRESS (2000/270) spectra, peaks of creatine, choline, N-acetylaspartate (NAA), and lactate resonances were integrated; for STEAM (2000/20) spectra, we measured the amplitude of the peaks at 3.2, 2.0, 1.3 and 0.9 ppm.RESULTSAn elevated lactate doublet was observed in the PRESS spectra. The choline/NAA ratio was 3.40. The amplitude ratios of the lipid pattern (0.9, 1.3 and 2.0 ppm) to choline were all below one.CONCLUSIONDespite the benign histology of the tumor, which generally lacks necrosis, a lactate signal was detected in all eight patients studied. A dominant lipid pattern was not observed.     

Intracranial tumors in children: small single-voxel proton MR spectroscopy using short- and long-echo sequences

We report preliminary experience using single-voxel, proton MR spectroscopy (MRS) employing small voxels of interest, in combination with short and long echo-time protocols, for the assessment of primary intracranial tumors in children. We examined 23 children with primary intracranial tumors detected by MRI, and 32 controls with similar ages, using MRS on a 1.5 T system. Localized single-voxel (3.7±1.3 cc) proton spectra were obtained with short-echo (2,000/18), stimulated-echo (STEAM) and long-echo (2,000/270) spin-echo (PRESS) protocols. All spectra were evaluated qualitatively; 10 tumor and 19 control spectra were processed for peak area quantification. Small voxels of interest were able to account for tissue heterogeneity. Combined acquisition of short- and long-echo spectra increased the number of detectable metabolites. The solid portion of all tumors exhibited reducedN-acetyl-aspartate (NAA), strong contribution from cholines (Cho) and inositols or glycine, minimal presence of total creatine (tCr), enhanced broad mobile lipid resonances and accumulated lactate. Calculated selected metabolite ratios of Cho/tCr and Cho/NAA were substantially increased from control values. The cystic portions of the masses showed only lipid and lactate peaks.     

Localized in vivo 1H NMR detection of neurotransmitter labeling in rat brain during infusion of [1-13C] D-glucose.

Resolved localized nuclear magnetic resonance (NMR) signals of 1H bound to 13C label in the carbon positions of glutamate C4, C3 and glutamine C4, C3, as well as in aspartate C3, lactate C3, alanine C3, gamma-aminobutyric acid C3, and glucose C1 were simultaneously observed in spectra obtained from rat brain in vivo. Time-resolved label incorporation was measured with a new adiabatic carbon editing and decoupling (ACED) single-voxel stimulated echo acquisition mode (STEAM) sequence. Adiabatic carbon broadband decoupling of 12 kHz bandwidth was achieved in vivo, which decoupled the entire 13C spectrum at 9.4 T. Resonances from N-acetyl-aspartate and creatine were also detected, consistent with natural-abundance 13C levels. These results emphasize the potential of 1H NMR for following complex biochemical pathways in localized areas of resting rat brain as well as during focal activation using infusions of 13C-labeled glucose.     

Cerebral glucose is detectable by localized proton NMR spectroscopy in normal rat brain in vivo.

This contribution reports the first direct and noninvasive observation of cerebral glucose in normal anesthetized rats (n = 16) using short-echo-time localized proton NMR spectroscopy (2.35 T, STEAM, TR = 6000 ms, TE = 20 ms, 125 microliters). In addition to resonances from N-acetyl aspartate (NAA), glutamate, total creatine, cholines, taurine, and myoinositol, all spectra exhibit strongly coupled resonances from glucose (3.43, 3.80 ppm) that are readily identifiable using model solutions. The observed level of cerebral glucose in fasted rats covered a range of 15-40% of that of NAA giving absolute concentrations of 1.1-2.8 mM when NAA is taken to be 7 mM. The arterial blood glucose concentration was 7.7 +/- 0.8 mM in the same group of animals.     

In vivo 1H NMR spectroscopy of the human brain at 7 T.

In vivo 1H NMR spectra from the human brain were measured at 7 T. Ultrashort echo-time STEAM was used to minimize J-modulation and signal attenuation caused by the shorter T2 of metabolites. Precise adjustment of higher-order shims, which was achieved with FASTMAP, was crucial to benefit from this high magnetic field. Sensitivity improvements were evident from single-shot spectra and from the direct detection of glucose at 5.23 ppm in 8-ml volumes. The linewidth of the creatine methyl resonance was at best 9 Hz. In spite of the increased linewidth of singlet resonances at 7 T, the ability to resolve overlapping multiplets of J-coupled spin systems, such as glutamine and glutamate, was substantially increased. Characteristic spectral patterns of metabolites, e.g., myo-inositol and taurine, were discernible in the in vivo spectra, which facilitated an unambiguous signal assignment.     

Localized proton NMR spectroscopy using stimulated echoes: applications to human skeletal muscle in vivo.

Localized proton NMR spectroscopy using stimulated echoes (STEAM) has been used to study metabolites in different proximal skeletal muscles of normal volunteers at rest. Single scan water-suppressed proton NMR spectra obtained at 1.5 and 2.0 T (Siemens Magnetom) from a 64-ml volume-of-interest (VOI) yield resonances due to triglycerides, phosphocreatine plus a minor contribution from creatine, and betaines comprising carnitine and choline-containing compounds. The observation of the pH-dependent resonances of carnosine required multiple acquisitions and echo times as short as 20 ms. T1 and T2 relaxation times of muscle metabolites were obtained by varying the repetition time and echo time of the STEAM sequence, respectively. Although rather long T2 values such as 180 ms for (phospho-) creatine correspond to natural resonance linewidths of only 2 Hz, the observed linewidths of typically 10-12 Hz are entirely determined by the short T2 relaxation times (25-30 ms) of the water protons used for shimming. The spectroscopic results from 24 muscle studies on 17 young male volunteers show remarkable intra- and interindividual differences in the absolute signal intensities of mobile lipids. Further metabolic variations were observed for the relative concentrations of betaines (by a factor of 2) and carnosine (by a factor of 3) when total creatine is assumed to be constant.     

Comparison of the quantification precision of human short echo time (1)H spectroscopy at 1.5 and 4.0 Tesla.

Precise quantification of human in vivo short echo time (1)H spectra remains problematic at clinical field strengths due to broad peak linewidths and low signal-to-noise ratio (SNR). In this study, multiple STEAM spectra (TE = 20 ms, volume = 8 cm(3)) were acquired in a single individual at 1.5 T and 4 T to compare quantification precision. Test-retest STEAM spectra (volume = 1.5 cm(3)) were also acquired from the anterior cingulate and thalamus of 10 individuals at 4.0 T. Metabolite levels were quantified using automated software that incorporated field strength-specific prior knowledge. With the distinct methods of data acquisition, processing, and fitting used in this study, peak height SNR increased approximately 80% while peak linewidth increased by approximately 50% in the 8 cm(3) volumes at 4.0 T compared to 1.5 T, resulting in an average increase in quantification precision of 39%. Metabolite levels from test-retest data (1.5 cm(3) voxels at 4.0 T) were quantified with similar inter- and intraindividual variability. Magn Reson Med 44:185-192, 2000. Published 2000 Wiley-Liss, Inc.     

Molecular self-diffusion of intracellular metabolites in rat brain in vivo Investigated by Localized Proton NMR Diffusion Spectroscopy

Molecular self-diffusion coefficients of water (0.75 ± 0.05), Nacetylaspartate (0.27 ± 0.04), creatines (0.27 ± 0.04), and cholines (0.28 ± 0.08) × 10^?5 cm² s^?1 were obtained from localized proton NMR spectra of rat brain in vivo using diffusion-weighted stimulated-echo (STEAM) sequences with a diffusion time of (Δ ? δ/3) = 17 ms.     

Reliable detection of macromolecules in single-volume 1H NMR spectra of the human brain.

In short echo time proton MR spectra of the brain, resonances from macromolecules are visible. The macromolecular resonances in the 0.5-2.0 ppm region can be affected by lipid contamination arising from fat-containing regions outside the selected volume of interest (VOI). This study demonstrates that considerable lipid contamination may remain in stimulated echo acquisition mode (STEAM) spectra even if the spoiling of unwanted coherences is sufficient and the VOI is placed 2 cm or more away from fat-containing regions. The observed contamination was attributed to residual remote out-of-volume excitation, although only very small out-of-slice ripples of less than 0.2% of the in-slice excitation were found in the calculated excitation profile of the RF pulses. Spatial presaturation of fat-containing regions led to a sufficient suppression of the contamination and enabled the detection of highly reproducible macromolecular resonances. Thus, in single-volume spectroscopy as well as in spectroscopic imaging (SI or CSI), the combination of volume selection and outer volume presaturation, each in three dimensions, is highly recommended to ensure accurate detection and reliable evaluation of even small pathological alterations in macromolecules, e.g., proteins or lipids, or other resonances in the 0.5-2.0 ppm region.     

Toward accurate quantification of metabolites,lipids, and macromolecules in HRMAS spectra of human brain tumor biopsies using LCModel

High‐resolution magic angle spinning (HRMAS) ¹H MR spectroscopy of biopsy samples provides detailed biochemical profiles that can be related to the lower‐resolution spectra obtained in vivo. Nevertheless, there is still significant overlap of many resonance peaks and contributions from broad lipid and macromolecule resonances that impede accurate quantification. We determined a minimum set of in vitro metabolite and simulated lipid and macromolecule resonances needed for LCModel analysis and quantification of brain tumor biopsy HRMAS spectra. We also demonstrate the quality of the LCModel fit for the four main brain tumor types (astrocytoma grade II, glioblastoma, metastasis, and meningioma). Our data suggest that when fitting resonances of coupled spins systems in high‐resolution spectra, interactions between metabolites and the macromolecular environment of the biopsy may cause small peak shifts not found in the solution spectra. However, LCModel is shown to provide a user‐independent method of analyzing HRMAS brain tumor spectra. Magn Reson Med, 2008. © 2008 Wiley‐Liss, Inc.     

Proton magnetic resonance spectroscopy with metabolite nulling reveals regional differences of macromolecules in normal human brain

PURPOSE: To quantify the macromolecular content in different anatomic brain regions and to evaluate an age dependency of the macromolecular concentrations. MATERIAL AND METHODS: A short echo time Stimulated Echo Acquisition Mode (STEAM) sequence was used without and with inversion recovery metabolite nulling in 8-12 healthy volunteers. Quantitation was achieved by an extended LCModel, and macromolecular resonances at 0.9, 1.4, 2.1, and 3.0 ppm were evaluated. RESULTS: In the cerebellum, the 1.4, 2.1, and 3.0 ppm resonances were highest compared to all other regions (P < 0.02); the 0.9 ppm resonance was significantly higher than that of pons (P < 0.01). In the motor cortex, the 0.9, 1.4, and 2.1 ppm resonances were higher than those of white matter and pons (P < 0.02). Pons and white matter did not differ significantly from each other. A significant correlation of the macromolecular concentrations with the age could not be found. CONCLUSION: There were higher macromolecular concentrations in the cerebellum and motor cortex than in pons or white matter. These were probably due to the higher portions of gray matter in these volumes of interest (VOIs) than in the other regions.     

In vivo 1H NMR spectroscopy of rat brain at 1 ms echo time.

Using optimized, asymmetric radiofrequency (RF) pulses for slice selection, the authors demonstrate that stimulated echo acquisition mode (STEAM) localization with ultra-short echo time (1 ms) is possible. Water suppression was designed to minimize sensitivity to B1 inhomogeneity using a combination of 7 variable power RF pulses with optimized relaxation delays (VAPOR). Residual water signal was well below the level of most observable metabolites. Contamination by the signals arising from outside the volume of interest was minimized by outer volume saturation using a series of hyperbolic secant RF pulses, resulting in a sharp volume definition. In conjunction with FASTMAP shimming (Gruetter Magn Reson Med 1993;29: 804-811), the short echo time of 1 msec resulted in highly resolved in vivo 1H nuclear magnetic resonance spectra. In rat brain the water linewidths of 11-13 Hz and metabolite singlet linewidths of 8-10 Hz were measured in 65 microl volumes. Very broad intense signals (delta v(1/2) > 1 kHz), as expected from membranes, for example, were not observed, suggesting that their proton T2 are well below 1 msec. The entire chemical shift range of 1H spectrum was observable, including resolved resonances from alanine, aspartate, choline group, creatine, GABA, glucose, glutamate, glutamine, myo-inositol, lactate, N-acetylaspartate, N-acetylaspartylglutamate, phosphocreatine, and taurine. At 9.4 T, peaks close to the water were observed, including the H-1 of alpha-D-glucose at 5.23 ppm and a tentative H-1 resonance of glycogen at 5.35 ppm.